本文選題：羊膜勻漿提取液 + 共聚焦顯微鏡�。� 參考：《南京大學(xué)》2013年碩士論文

【摘要】：目的： 觀(guān)察人羊膜勻漿提取液對(duì)大鼠角膜堿燒傷后角膜超微結(jié)構(gòu)的影響,探討羊膜勻漿提取液應(yīng)用于角膜堿燒傷后的臨床價(jià)值及作用機(jī)理。 方法： 健康無(wú)眼疾的SD大鼠40只(40眼)隨機(jī)分為實(shí)驗(yàn)組和對(duì)照組,每組20只(20只眼)。相同條件下,所有大鼠使用lmol/L的NaOH溶液建立右眼角膜堿燒傷模型。損傷后實(shí)驗(yàn)組立即行人羊膜勻漿提取液點(diǎn)眼,每日四次,對(duì)照組用PBS液點(diǎn)眼,每日四次。堿燒傷后每日在裂隙燈顯微鏡下觀(guān)察大鼠眼表的總體情況,例如眼結(jié)膜充血,角膜透明性,以及新生血管生長(zhǎng)情況。進(jìn)行眼前節(jié)照相,通過(guò)圖像分析系統(tǒng)記錄兩組大鼠角膜上皮愈合,以及角膜新生血管生長(zhǎng)情況。隔日行共聚焦顯微鏡動(dòng)態(tài)觀(guān)察大鼠角膜超微結(jié)構(gòu)的變化,分析角膜中炎癥細(xì)胞、成纖維細(xì)胞以及內(nèi)皮細(xì)胞的數(shù)量及形態(tài)改變。于燒傷后3d、7d、14d、28d每組隨機(jī)處死3只試驗(yàn)大鼠,取大鼠眼球進(jìn)行HE染色,行組織學(xué)染色觀(guān)察角膜各層超微結(jié)構(gòu)的變化。 結(jié)果： 堿燒傷后第1d至第7d,實(shí)驗(yàn)組角膜上皮愈合明顯快于對(duì)照組(P0.01)。實(shí)驗(yàn)組角膜新生血管出現(xiàn)平均時(shí)間為(4.8±0.6)d,晚于對(duì)照組(3.1±0.5)d,(P0.01),對(duì)照組最大新生血管面積為(48.72±2.12)mm2,實(shí)驗(yàn)組為(35.12±1.77)mm2,各個(gè)時(shí)間點(diǎn)兩組之間角膜新生血管面積的差異均有統(tǒng)計(jì)學(xué)意義(各p0.01)。堿燒傷后9d,對(duì)照組炎癥細(xì)胞數(shù)量最多為(3164±306)個(gè)/mm2,實(shí)驗(yàn)組為(2830±254)個(gè)/mm2,兩者之間差異有統(tǒng)計(jì)學(xué)意義(P0.01)；堿燒傷后11d,對(duì)照組成纖維細(xì)胞數(shù)量為(349±105)個(gè)/mm2,實(shí)驗(yàn)組為(248±84)個(gè)/mm2,兩者之間差異有統(tǒng)計(jì)學(xué)意義(P0.01)；堿燒傷后28d,對(duì)照組內(nèi)皮細(xì)胞計(jì)數(shù)為(1837±59)個(gè)/mm2,對(duì)照組內(nèi)皮細(xì)胞計(jì)數(shù)為(2063±54)個(gè)/mm2,兩者之間差異有統(tǒng)計(jì)學(xué)意義(P0.01)。HE染色提示實(shí)驗(yàn)組炎癥細(xì)胞、成纖維細(xì)胞、角膜新生血管數(shù)量較少,基質(zhì)層纖維結(jié)構(gòu)排列較為規(guī)則。結(jié)論： 新鮮人羊膜勻漿提取液可以有效促進(jìn)大鼠堿燒傷后角膜上皮細(xì)胞的增殖,抑制炎癥細(xì)胞浸潤(rùn),減少角膜新生血管的形成,抑制成纖維細(xì)胞的激活和增殖,減少角膜瘢痕的形成,有利于堿燒傷后大鼠角膜的透明愈合。
[Abstract]:Objective: To observe the effect of human amniotic membrane homogenate extract on the ultrastructure of cornea after alkali burn of rat cornea, and to explore the clinical value and mechanism of application of amniotic membrane homogenate extract to corneal alkali burn. Methods: Forty SD rats without eye disease were randomly divided into experimental group and control group with 20 eyes in each group. Under the same conditions, all the rats used NaOH solution of lmol/L to establish the right eye corneal alkali burn model. After injury, the experimental group was treated with amniotic membrane homogenate extract, four times a day, while the control group was treated with PBS solution four times a day. After alkali burn, we observed the overall condition of eye surface under slit lamp microscope, such as conjunctival hyperemia, corneal transparency, and neovascularization. The corneal epithelium healing and corneal neovascularization were recorded by image analysis system. The changes of corneal ultrastructure were observed by confocal microscope on the following day, and the number and morphology of inflammatory cells, fibroblasts and endothelial cells in the cornea were analyzed. Three rats in each group were randomly killed on the 3rd day, 7th day, 14d and 28d after burn. The eyeballs of the rats were stained with HE, and the ultrastructure of each layer of cornea was observed by histological staining. Results: From the first day to the 7th day after alkali burn, the corneal epithelium healed faster in the experimental group than that in the control group (P 0.01). The mean time of corneal neovascularization in the experimental group was 4.8 鹵0.6 days, which was later than that in the control group (3.1 鹵0.5), the maximum neovascularization area in the control group was 48.72 鹵2.12mm2, and in the experimental group was 35.12 鹵1.77m2.There was significant difference in the corneal neovascularization area between the two groups at each time point (p0.01). At 9 days after alkali burn, the number of inflammatory cells in the control group was 3164 鹵306 / m2, and in the experimental group was 2830 鹵254 / m2, the difference was statistically significant (P 0.01); at 11 days after alkali burn, the number of fibroblasts in the control group was 349 鹵105 / mm2, and in the experimental group was 248 鹵84 / mm2, between the two groups. The number of endothelial cells in the control group was 1837 鹵59 / m2 and that in the control group was 2063 鹵54 / mm2 on the 28th day after alkali burn. The difference between the two groups was statistically significant (P 0.01) and HE staining indicated that the inflammatory cells were found in the experimental group. The number of corneal neovascularization in fibroblasts was less and the fibrous structure of stroma was regular. Conclusion: Fresh human amniotic membrane homogenate extract can effectively promote the proliferation of corneal epithelial cells, inhibit the infiltration of inflammatory cells, reduce the formation of corneal neovascularization, inhibit the activation and proliferation of fibroblasts, and reduce the formation of corneal scar. It is beneficial to the transparent healing of cornea after alkali burn in rats.
【學(xué)位授予單位】：南京大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R779.13

【參考文獻(xiàn)】

相關(guān)期刊論文 前3條

1 李琦;席興華;張艷彬;;人羊膜勻漿提取液抑制大鼠角膜堿燒傷后新生血管生成[J];國(guó)際眼科雜志;2007年05期

2 高髻云;張偉;;羊膜勻漿液對(duì)兔小梁切除術(shù)后濾過(guò)泡及TGFβ1、CTGF表達(dá)的影響[J];山東大學(xué)學(xué)報(bào)(醫(yī)學(xué)版);2008年02期

3 廖瓊,劉翔;羊膜勻漿上清液對(duì)兔角膜上皮細(xì)胞增殖的影響[J];眼科新進(jìn)展;2004年04期

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本文編號(hào)：1965664